This study used Langmuir-Schaefer (LS) method to produce thermo-responsive poly(N-isopropylacrylamide) (PIPAAm) modified surface. Block copolymer composed of polystyrene (PSt) and PIPAAm was synthesized by RAFT polymerization. PSt-block-PIPAAm (St-IP) with various chemical compositions was dropped on an air-water interface and formed Langmuir film by compression. Then, the Langmuir film changing a density was transferred on a hydrophobic modified glass substrate to produce St-IP transferred surface (St-IP LS surface). From the observation of atomic force microscope images, a nanostructure was observed on the transference of Langmuir films. Cell adhesion and detachment were also evaluated on the LS surfaces in response to temperature. Cell adhesion on LS surfaces at 37 °C was controlled by changing the chemical compositions and densities. After reducing temperature to 20 °C, adhering cells rapidly detached themselves with lower Am and higher composition of PIPAAm. Our method should be proved novel insights for investigating cell adhesion and detachment on thermo-responsive surfaces.